Abstract
NORFA, the first lincRNA associated with sow fertility, has been shown to control granulosa cell (GC) functions and follicular atresia. However, the underlying mechanism is not fully understood. In this study, RNA-seq was performed and we noticed that inhibition of NORFA led to dramatic transcriptomic alterations in porcine GCs. A total of 1,272 differentially expressed transcripts were identified, including 1167 DEmRNAs and 105 DEmiRNAs. Furthermore, protein–protein interaction, gene-pathway function, and TF–miRNA–mRNA regulatory networks were established and yielded four regulatory modules with multiple hub genes, such as AR, ATG5, BAK1, CENPE, NR5A1, NFIX, WNT5B, ssc-miR-27b, and ssc-miR-126. Functional assessment showed that these hub DEGs were mainly enriched in TGF-β, PI3K-Akt, FoxO, Wnt, MAPK, and ubiquitin pathways that are essential for GC states (apoptosis and proliferation) and functions (hormone secretion). In vitro, we also found that knockdown of NORFA in porcine GCs significantly induced cell apoptosis, impaired cell viability, and suppressed 17β-estradiol (E2) synthesis. Notably, four candidate genes for sow reproductive traits (INHBA, NCOA1, TGFβ-1, and TGFBR2) were also identified as potential targets of NORFA. These findings present a panoramic view of the transcriptome in NORFA-reduced GCs, highlighting that NORFA, a candidate lincRNA for sow fertility, is crucial for the normal states and functions of GCs.
Highlights
In mammalian ovaries, follicular development is a crucial biological process for follicle maturation, ovulation, and female fertility (Botta et al, 2017)
A high-throughput RNA sequencing strategy was designed to identify the crucial RNA molecules and core pathways involved in the responses of porcine granulosa cell (GC) to the knockdown of NORFA (Figure 1)
Porcine GCs were cultured for 48 h and transfected with siNORFA as Identification and Characterization of the DEmRNAs in NORFA-Inhibited Porcine GCs
Summary
Follicular development is a crucial biological process for follicle maturation, ovulation, and female fertility (Botta et al, 2017). As the main threat to female ovarian function, severe follicular atresia leads to follicular development arrest, premature follicles failure (PFF), and even infertility (Meng et al, 2017; Yeung et al, 2017). Increasing evidence has suggested that GC apoptosis and non-apoptotic programmed cell death are the main causes of follicular. In the first two decades of this century, a bunch of microRNAs (miRNAs) and circRNAs have been identified by using high-throughput sequencing technology and further proved to be involved in the regulation of follicular development, especially for GC states and functions (Du et al, 2016, 2018; Guo et al, 2019). The roles of long noncoding RNAs (lncRNAs) in follicular atresia and GC apoptosis have not been fully characterized
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call